酸提和酶解两步法连续提取鳙鱼鱼鳞胶原蛋白工艺研究
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摘要
采用柠檬酸提和木瓜蛋白酶解两步法从鳙鱼鱼鳞中连续提取胶原蛋白。采取正交实验方法对两步法工艺条件进行了优化,并与单一酸提和酶解工艺进行了比较。对酸提、酶解和酸提+酶解两步法制备的胶原蛋白样品进行了紫外和红外表征。结果表明,两步法中酸提最佳工艺条件为提取时间24 h、液固比15 mL/g、酸浓度0.5 mol/L、提取温度28℃,提取率9.25%;酶解最佳工艺条件为提取时间36 h、液固比20 mL/g、酶浓度2.0%、提取温度28℃,提取率56.23%。两步法总提取率65.48%,分别是单一酸提和酶解提取率的7.1倍和2.1倍。两步法胶原蛋白样品的紫外和红外谱图上的吸收峰位置与单一酸提和酶解的十分相似,均符合Ⅰ型胶原蛋白的基本特征。鱼鳞经一步酸提后发生溶胀,部分胶原交联键被打开,有利于下步酶解过程中酶分子与底物接触几率的提高,从而大幅度提高胶原蛋白的提取率,使酶解(两步法)的提取率(56.23%)比单一酶解法的提取率(31.20%)提高了80%,是一种高效的鱼鳞胶原蛋白的提取方法。
Two-step method of citric acid extraction and papain hydrolysis was used to extract collagen from bighead fish scales. The two-step method was optimized by orthogonal experiment and compared with single acid extraction and enzymatic hydrolysis. The collagen samples prepared by single acid extraction, single enzymatic hydrolysis and two-step method of acid extraction plus enzymatic hydrolysis were characterized by UV and IR. Results showed that in the two-step method, the optimum conditions of the acid extraction were: extracting time 24 h, liquid-solid ratio 15 mL/g, acid concentration 0.5 mol/L and extraction temperature 28 ℃, under which, the extraction rate was 9.25%. The optimum conditions of enzymatic hydrolysis were: extracting time 36 h, liquid-solid ratio 20 mL/g, enzyme concentration 2.0%,extraction temperature 28 ℃, under which the extraction rate was 56.23%. The total extraction rate of the two-step method was 65.48%, which was 7.1 times and 2.1 times of that of single-acid extraction and single enzymatic extraction, respectively. The absorption peaks of the collagen sample by twostep method appeared on the UV and IR spectra were very similar to those by single acid extraction and enzymatic hydrolysis, and were consistent with the basic characteristics of type Ⅰ collagen. The fish scales were swelled and part of the collagen cross-linking key was opened after acid extraction, which were conductive to increase the contact chances of enzyme molecules with substrate, greatly improved the extraction rate in the next step of enzymatic hydrolysis. The extraction rate of enzymatic extraction by the two-step method(56.23%)was increased by 80% compared with that of single enzyme hydrolysis(31.20%), meaning that the two-step method was a kind of very effective one for collagen extraction from fish scales.
Two-step method of citric acid extraction and papain hydrolysis was used to extract collagen from bighead fish scales. The two-step method was optimized by orthogonal experiment and compared with single acid extraction and enzymatic hydrolysis. The collagen samples prepared by single acid extraction, single enzymatic hydrolysis and two-step method of acid extraction plus enzymatic hydrolysis were characterized by UV and IR. Results showed that in the two-step method, the optimum conditions of the acid extraction were: extracting time 24 h, liquid-solid ratio 15 mL/g, acid concentration 0.5 mol/L and extraction temperature 28 ℃, under which, the extraction rate was 9.25%. The optimum conditions of enzymatic hydrolysis were: extracting time 36 h, liquid-solid ratio 20 mL/g, enzyme concentration 2.0%,extraction temperature 28 ℃, under which the extraction rate was 56.23%. The total extraction rate of the two-step method was 65.48%, which was 7.1 times and 2.1 times of that of single-acid extraction and single enzymatic extraction, respectively. The absorption peaks of the collagen sample by twostep method appeared on the UV and IR spectra were very similar to those by single acid extraction and enzymatic hydrolysis, and were consistent with the basic characteristics of type Ⅰ collagen. The fish scales were swelled and part of the collagen cross-linking key was opened after acid extraction, which were conductive to increase the contact chances of enzyme molecules with substrate, greatly improved the extraction rate in the next step of enzymatic hydrolysis. The extraction rate of enzymatic extraction by the two-step method(56.23%)was increased by 80% compared with that of single enzyme hydrolysis(31.20%), meaning that the two-step method was a kind of very effective one for collagen extraction from fish scales.
引文
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[2]Zhong-Rui Li,Bi Wang,Chang-Feng Chi,et al.Isolation and characterization of acid soluble collagens and pepsing soluble collagens from the skin and bone of Spanish mackerel(Scomberomorous niphonius)[J].Food Hydrocolloids,2013,31:103-113.
[3]Hideki Mori,Yurie Tone,Kouske Shimizu,et al.Studies on fish scale collagen of pacific saury(Cololabis saira)[J].Materials Science&Engineering C,2013,33:174-181.
[4]周玉惠.2种提取草鱼鱼鳞胶原蛋白工艺比较[J].山地农业生物学报,2013,32(3):254-257.
[5]胡建平.鱼鳞胶原蛋白的酸法提取及性质研究[J].食品科技,2012,(11):141-148.
[6]杨平,刘影,公丽艳,等.酸法和酶法提取草鱼鱼鳞胶原蛋白的特性分析[J].食品工业,2018,39(7):129-132.
[7]卢昭.草鱼鱼鳞胶原蛋白提取分离工艺的研究[D].合肥:安徽农业大学,2013.
[8]佘广麟,蔡阳伦,何志强,等.鱼鳞胶原蛋白的提取方法研究[J].广东化工,2015,42(16):57-58.
[9]酶解法提取草鱼鱼鳞胶原蛋白的工艺研究[J].新余学院学报,2014,19(2):10-12.
[10]王希搏.鱼鳞有机酸钙、胶原蛋白及胶原多肽联产工艺与工厂设计[D].广州:华中农业大学,2013.
[11]王春,周莉,周天,等.木瓜蛋白酶提取罗非鱼鱼鳞胶原蛋白的研究[J].安徽农业科学,2013,41(3):1291-1292.
[12]胡杨,王希搏,熊善柏,等.一种以鱼鳞为原料的多产物联产工艺的建立与优化[J].食品工业科技,2016,37(24):197-202.
[13]罗灿.草鱼鱼鳞胶原蛋白与角蛋白的分离纯化及鉴定[D].长沙:湖南农业大学,2015.
[14]梁学世.罗非鱼鱼鳞胶原蛋白提取条件优化及其功能膜制备的研究[D].广州:华南师范大学,2013.
[15]聂小宝,潘洪民,程丽林,等.鲫鱼鱼鳞胶原蛋白提取工艺的研究[J].山东农业科学,2012,44(1):109-111.
[16]LIU D S,LIANG L,REGENESTEIN J M,et al.Extraction and characterization of pepsin-solubilised collagen from fins,scales,bones and swim bladders of bighead carp(Hypophthalmichthys nobilis)[J].Food Chemistry,2012,133(4):1441-1448.
[17]冯文婕,赵粼,阙斐.酸酶复合法优化鱼鳞胶原蛋白的提取工艺[J].湖北农业科学,2016,55(5):1242-1246.
[18]孔丽丽,巩凡,Viktoriia Plavan,等.酸酶复合法提取鲢鱼鱼鳞胶原蛋白的工艺研究[J].皮革科学与工程,2014,24(4):11-15.
[19]陈铁壁,刘冬敏,鹿康,等.草鱼鱼鳞胶原蛋白酸酶分步提取工艺研究[J].食品与机械,2016,32(8):163-166.
[20]李彦春,程宝缄,立强.胶原蛋白的应用[J].皮革化工,2001,19(3):10-14.